Figure 1. Design and activity analysis of native FGF2 and its
variants. (A ) Based on the crystal structure of FGF2 (PDB ID:
1FQ9), FGF2-FGFR1-heparin complex structure (left panel), close-up view
of two surface-exposed residues, Lys129 and Phe17, near the heparin- and
receptor-binding regions, respectively, and two surface-exposed
cysteines (Cys69 and Cys87) on FGF2 (right panel). (B )
Schematic representation of two FGF2 variants, including
FGF2C69A (1 ) and FGF2C87A(2 ). The residues labeled in green indicate mutation sites, and
the residues marked in red are single conjugation sites for each FGF2
variant. (C ) Immunoblotting analysis of p-ERK1/2 and ERK1/2
levels in cell lysates of HUVECs after treatment with
compounds 1 , 2 (10
ng·mL-1). ERK was used as a loading control (n = 3).
(E ) Schematic representation of another two synthetic FGF2
variants, including FGF2C69A/C87A/F17C (3 )
and FGF2C69A/C87A/K129C (4 ). (F )
Immunoblotting analysis of p-ERK1/2 and ERK1/2 levels in cell lysates of
HUVECs after treatment with compound 3 , 4 (10
ng·mL-1) (n = 3). (D, G ) Quantitative
analysis of p-ERK1/2 in the indicated groups. Data are displayed as
means ± SEM. **P < 0.01,***P < 0.001 vs. control group;#P < 0.05 vs. FGF2 group; n.s., no
significance.